Figure 7.

hTRMT2A has no impact on rRNA methylation, but affects translation fidelity. (A) Barplot of rRNA MS analysis of m⁵C in 28S and 18S rRNA shows two modifications in 28S, as reported earlier. (B) Barplot of rRNA MS analysis of m⁵U in 28S and 18S rRNA shows very low modification levels that do not change upon hTRMT2A KD. rRNA was obtained from ribosomes that were isolated from hTRMT2A WT and KD1 and KD2 cells. Data represents the mean ± SD of three biological replicates. (C) Translation fidelity assay with dual luciferase WT, E145Q and D120N mutants, and their respective luminescence readout indicated over the graph. Luciferase plasmids were transfected into hTRMT2A WT, KD1 and KD2. For the E145Q luciferase mutant, the ratio in KD1 and KD2 was significantly increased over that in the WT, indicating a higher translation error rate. WT and the D120N luciferase mutant did not show significant changes. Data represent the mean ± SD of six biological replicates in technical triplicates. Significance was tested with Wilcoxon test for non-normally distributed values with P > 0.05 = ns, P < 0.05 = significant.